目的探讨丝裂原和应激激活蛋白激酶1（mitogen-and stress-activated protein kinase1，MSK1）及cAMP反应元件结合蛋白（cAMP response element binding protein，CREB）在氯胺酮保护小鼠缺血脑组织中的作用。方法80只健康成年雄性BALB...目的探讨丝裂原和应激激活蛋白激酶1（mitogen-and stress-activated protein kinase1，MSK1）及cAMP反应元件结合蛋白（cAMP response element binding protein，CREB）在氯胺酮保护小鼠缺血脑组织中的作用。方法80只健康成年雄性BALB／c小鼠按完全随机法分为5组[每组16只，其中包括行为学检测、氯化三苯基四氮唑（triphenyltetrazolium chloride，TTc）染色10只，Westernblot检测6只]：假手术组，大脑中动脉阻塞（middle cerebral artery occlusion，MCAO）＋生理盐水组，MCAO＋25、50、100mg／kg氯胺酮组。利用小鼠MCAO模型，结合神经行为学评分、TTC染色和Western blot等技术，检测小鼠脑皮质不同缺血区域内MSK1和CREB磷酸化水平和蛋白总量的表达，观察不同剂量氯胺酮对小鼠脑缺血损伤的影响。结果与MCAO＋生理盐水组比较，MCAO＋25mg/kg氯胺酮组小鼠脑缺血后神经行为学表现明显改善[（8．2±0．4）分比（6．7±0．3）分]，脑缺血皮质梗死体积减少[（28．3±2．0）％比（21．0±2．2）％]，水肿率降低[（10．6±0．4）％比（6．9±0．9）％]，缺血半影区内MSK1[（52．3＋7．7）％比（81．1±6．9）％]和CREB[（56．6±5．9）％比（78．6±7．1）％]磷酸化水平明显增加（P〈0．05）；而MSK1和CREB总蛋白表达量在缺血核心区和半影区内均无明显改变。MCAO＋50、100mg／kg氯胺酮组小鼠行为学表现，缺血皮质梗死体积，水肿率，及缺血半影区MSK1、CREB磷酸化水平无明显改变。结论小剂量氯胺酮降低小鼠缺血性脑损伤可能与缺血皮质半影区MSK1及其底物CREB的磷酸化水平增高有关。展开更多
The Keap1–Nrf2–ARE pathway is an important antioxidant defense mechanism that protects cells from oxidative stress and the Keap1–Nrf2 protein–protein interaction(PPI) has become an important drug target to upregul...The Keap1–Nrf2–ARE pathway is an important antioxidant defense mechanism that protects cells from oxidative stress and the Keap1–Nrf2 protein–protein interaction(PPI) has become an important drug target to upregulate the expression of ARE-controlled cytoprotective oxidative stress response enzymes in the development of therapeutic and preventive agents for a number of diseases and conditions. However, most known Nrf2 activators/ARE inducers are indirect inhibitors of Keap1–Nrf2PPI and they are electrophilic species that act by modifying the sulfhydryl groups of Keap1's cysteine residues. The electrophilicity of these indirect inhibitors may cause "off-target" side effects by reacting with cysteine residues of other important cellular proteins. Efforts have recently been focused on the development of direct inhibitors of Keap1–Nrf2 PPI. This article reviews these recent research efforts including the development of high throughput screening assays, the discovery of peptide and small molecule direct inhibitors, and the biophysical characterization of the binding of these inhibitors to the target Keap1 Kelch domain protein. These non-covalent direct inhibitors of Keap1–Nrf2 PPI could potentially be developed into effective therapeutic or preventive agents for a variety of diseases and conditions.展开更多
基金the financial support of Grants CA133791, CA125868, and MH093197 from the National Institutes of Health, United States
文摘The Keap1–Nrf2–ARE pathway is an important antioxidant defense mechanism that protects cells from oxidative stress and the Keap1–Nrf2 protein–protein interaction(PPI) has become an important drug target to upregulate the expression of ARE-controlled cytoprotective oxidative stress response enzymes in the development of therapeutic and preventive agents for a number of diseases and conditions. However, most known Nrf2 activators/ARE inducers are indirect inhibitors of Keap1–Nrf2PPI and they are electrophilic species that act by modifying the sulfhydryl groups of Keap1's cysteine residues. The electrophilicity of these indirect inhibitors may cause "off-target" side effects by reacting with cysteine residues of other important cellular proteins. Efforts have recently been focused on the development of direct inhibitors of Keap1–Nrf2 PPI. This article reviews these recent research efforts including the development of high throughput screening assays, the discovery of peptide and small molecule direct inhibitors, and the biophysical characterization of the binding of these inhibitors to the target Keap1 Kelch domain protein. These non-covalent direct inhibitors of Keap1–Nrf2 PPI could potentially be developed into effective therapeutic or preventive agents for a variety of diseases and conditions.
